Wei Jun-ji, Zeng Li-fen, Fan Xiao-tong, Wang Yu, Ma Wen-bin, Li Gui-lin, Dou Wan-chen, Zhang Zhen-xing, Li Shi-fang, Feng Ming, Han Qin, Li Zhao-jian, Zhang Zi-heng, Kang Jun, Kong Yan-guo, Wang Ren-zhi, Zhao Chun-hua
Department of Neurosurgery, Peking Union Medical College Hospital, Chinese Academy of Medical Science & Peking Union Medical College, Beijing 100730, China.
Zhonghua Yi Xue Za Zhi. 2007 Jan 16;87(3):184-9.
To investigate the effects of treatment of stroke in rats with bone marrow mesenchymal stem cells (BMSCs) and mechanism thereof.
Bone marrow of a healthy volunteer was collected and the BMSCs were separated with density gradient centrifugation. The hBMSC were cultivated and harvested until the third passage. A number of adult male Sprague-Dawley rats received corresponding behavioral training before surgery and underwent transient middle cerebral arterial occlusion (MCAO) for 2 hours. Sixty of them showing the scores of 6 approximately 12 according to the modified neurological severity score system were randomly divided into 2 groups: treatment group (n = 48, injected into the cortex around the ischemic areas with hBMSCs 3x10(5)/15 microl) and control group (n = 12, injected with D-Hanks solution 15 microl 24 hours after the establishment of MCAO models. Morris water maze test, Rotarod test and adhesive-removal test were performed since the 4th day to the 32 day after transplantation once every 3 days. 1, 2, 3, and 4 weeks after the transplantation 12 rats from each group were killed randomly to take out their brains. Immunofluorescence was used to identify the migration, survival and differentiation of the hBMSC.
A large number of hBMSC could be seen within 2 weeks after transplantation. The number of hBMSC decreased since the 21st day after transplantation and few cells could be found at the end of 1 month after. No definite evidence supported the differentiation of neural cells derived from the hBMSCs during the whole process. Morris water maze test showed that the mean escape time 1 week after transplantation of the treatment group was (69 +/- 10) s, significantly shorter than that of the control group [(120 +/- 0) s, P < 0.05] The significant difference persisted until the 4(th) week (P > 0.05). Rotarod test with the speed of 10 r/min showed that the mean latency period 10 days after transplantation of the treatment group was (167 +/- 18) s, significantly longer than that of the control group [(37 +/- 19) s, P < 0.05]. The significant difference persisted until the experimental terminal. The adhesive-removal test showed that the mean latency period 13 days after transplantation of the treatment group was (33 +/- 8) s, significant shorter than that of the control group [(84 +/- 13) s, P < 0.05]. The significant difference persisted until the experimental terminal.
Injection of hBMSCs into brain cortex improves neurological functional recovery after stroke. The transplanted cells can migrate and survive for a certain period, but no hBMSC express proteins phenotype of neural cells.
探讨骨髓间充质干细胞（BMSCs）治疗大鼠脑卒中的效果及其机制。
采集健康志愿者的骨髓，采用密度梯度离心法分离BMSCs。培养并收获人BMSCs至第3代。若干成年雄性Sprague-Dawley大鼠在手术前接受相应行为训练，然后进行大脑中动脉短暂闭塞（MCAO）2小时。其中60只根据改良神经功能缺损评分系统评分为6至12分的大鼠被随机分为2组：治疗组（n = 48，于缺血区周围皮质注射3×10(5)/15微升人BMSCs）和对照组（n = 12，在MCAO模型建立后24小时注射15微升D-Hanks液）。自移植后第4天至第32天，每3天进行一次Morris水迷宫试验、转棒试验和黏贴去除试验。移植后1、2、3和4周，每组随机处死12只大鼠并取脑。采用免疫荧光法鉴定人BMSCs的迁移、存活及分化情况。
移植后2周内可见大量人BMSCs。移植后第21天起人BMSCs数量减少，移植后1个月末几乎未见细胞。整个过程中无确切证据支持人BMSCs分化为神经细胞。Morris水迷宫试验显示，治疗组移植后1周的平均逃避潜伏期为(69±10)秒，显著短于对照组[(120±0)秒，P<0.05]。该显著差异持续至第4周（P>0.05）。转棒试验中转速为10转/分钟时，治疗组移植后10天的平均潜伏期为(167±18)秒，显著长于对照组[(37±19)秒，P<0.05]。该显著差异持续至实验结束。黏贴去除试验显示，治疗组移植后13天的平均潜伏期为(33±8)秒，显著短于对照组[(84±13)秒，P<0.05]。该显著差异持续至实验结束。
向脑皮质注射人BMSCs可改善脑卒中后神经功能恢复。移植细胞可迁移并存活一段时间，但无人BMSCs表达神经细胞蛋白表型。
